The effect of re-release of Oryctes rhinoceros baculovirus in the biological control of rhinoceros beetles in Western Samoa

The Rhinoceros Beetle Project in Western Samoa has developed and successfully applied biological methods to control the rhinoceros beetle, a serious pest of coconut palms, by using two specific pathogens, a baculovirus (Family Baculoviridae), and an entomopathogenic fungus, Metarhizium anisopliae. The application of virus particularly has markedly suppressed the beetle population and helped revive the copra industry. The virus disease had established itself in the wild beetle population several years after its introduction at a level between 30 and 50%. At the same time an increase in beetle numbers and damage to palm trees was experienced. Therefore, a continuous release of virus into beetle-infested areas was proposed. It was argued that, considering the relatively high level of “natural” virus incidence, further releases of virus into the population would be futile. In a combined research and control program, virus was again re-released into the wild beetle population which was already virus infected. The results show that through re-release the virus level can be raised and the number of beetles and consequently the damage can be reduced. The techniques of the control methods are described. The virus release is very easy and cheap; it requires no chemicals, no special equipment, and it is particularly recommended in situations where breeding places are inaccessible or other methods such as plantation sanitation are either impossible or economically impractical. Above all, the methods are absolutely safe from the standpoint of environmental protection.     

The Oryctes virus: its detection, identification, and implementation in biological control of the coconut palm rhinoceros beetle, Oryctes rhinoceros (Coleoptera: Scarabaeidae)

In view of the increasing and devastating damage by rhinoceros beetle (Oryctes rhinoceros) to coconut palms in the middle of last century, many efforts were made to find an efficient natural control factor against this pest, which could not be controlled by pesticides. The basic procedures of these monitoring programmes are outlined together with the final detection of a virus disease in oil palm estates in Malaysia in 1963. In extensive laboratory studies, the virus was isolated and identified as the first non-occluded, rod-shaped insect virus, morphologically resembling the baculoviruses. Infection experiments clarified the pathology, histopathology, and virulence of the virus and demonstrated that the virus was extremely virulent to larvae after peroral application. These findings encouraged the first pilot release of virus in 1967 in coconut plantations of Western Samoa where breeding sites were contaminated with virus. Surprisingly, the virus became established in the Samoan rhinoceros beetle populations and spread autonomously throughout the Western Samoan islands. As a consequence, there was a drastic decline of the beetle populations followed by a conspicuous recovery of the badly damaged coconut stands. This unexpected phenomenon could only be explained after it was shown that the adult beetle itself is a very active virus vector and thus was responsible for the efficient autodissemination of the virus. The functioning of the beetle as a 'flying virus factory' is due to the unique cytopathic process developing in the midgut after peroral virus infection. Pathological details of this process are presented. Because of the long-term persistence of the virus in the populations, rhinoceros beetle control is maintained. Incorporation of virus into integrated control measures and successful virus releases in many other countries are recorded.     

The epizootiology of two pathogens of the coconut palm rhinoceros beetle

Two diseases of Oryctes rhinoceros caused by Metarrhizium anisopliae and a virus were introduced from Western Samoa into Tongatapu, Kingdom of Tonga, in 1969 and 1970, respectively. The fungus remained at a low incidence and did not significantly affect the beetle population. The virus disease developed to epizootic levels within 5 months and spread across the island in 15 months. The beetle population was monitored through palm damage surveys, which confirmed that the population was considerably reduced by the disease.The greater impact of the virus was attributed to its much higher power of dispersal through infected beetles.The experiment supports the view that the reduction in beetle numbers recorded previously in Western Samoa was caused by a virus epizootic.     

Microbial pathogens of the coconut pest Oryctes rhinoceros: influence of weather factors on their infectivity and study of their coincidental ecology in Kerala,India

The rhinoceros beetle, Oryctes rhinoceros L., is an economically important pest of the coconut palm. Management of this pest has been accomplished using microbial agents viz., Oryctes virus (OrV) and an entomofungal pathogen Metarhizium anisopliae. Recently an opportunistic bacterial pathogen Pseudomonas alcaligenes has also been noticed to cause septicaemia in the grubs when under stress. To unravel the influence of abiotic weather factors and the interactions amongst these microbial pathogens, a 3 year study was conducted from September 1996 to August 1999 in three of the southern districts of Kerala, India. Of the 6627 grubs and 307 adults collected from various breeding sites of the pest, 5% of the grubs and 22% of the adults had natural virus infection, 3% larvae died of M. anisopliae mycosis and 20% larvae succumbed to bacterial septicaemia. Oryctes virus infection in grubs and adults was negatively correlated to minimum temperature (correlation co-efficient, r = –0.4, and –0.6 respectively, sample size, n = 0). Increase in relative humidity increased the fungal activity (r = 0.8) whereas, maximum temperature had a negative impact (r = –0.7). Occurrence of virus infection in grubs and adults was positively correlated (r = 0.6), supporting the contention of active transmission of the virus pathogen between these two stages. The bacterial septicaemia in the grubs was marginally correlated with virus infection and P. alcaligenes undermined the efficiency of the virus pathogen.     

Ecology of baculovirus-infected and healthy adults of Oryctes rhinoceros (Coleoptera: Scarabaeidae) on coconut palms in the Philippines

Abstract.

• 1 Males and females of Oryctes rhinoceros (L.), a serious pest of coconut palms, congregate in decaying coconut trunks to mate, oviposit and prepare the wood for the young larvae. A baculovirus disease is often transmitted from beetle to beetle in this environment and transmission to and from larvae probably also occurs.
• 2 Young beetles are mainly found feeding on palms, where they also frequently contract the virus disease, presumably through mating with diseased partners.
• 3 Monitoring beetle populations in the Philippines with attractant traps and by collections from palms showed seasonal fluctuations which were often inversely correlated with the incidence of the virus disease. Disease prevalence matched closely the proportion of females in the population. Possible explanations for this correlation and for the seasonality of the disease prevalence are outlined.

     

The incidence and use of Oryctes virus for control of rhinoceros beetle in oil palm plantations in Malaysia

The rhinoceros beetle, Oryctes rhinoceros, has emerged as a serious pest of oil palm since the prohibition of burning as a method for maintaining estate hygiene in the 1990s. The abundance of beetles is surprising given that the Malay peninsula was the site of first discovery of the Oryctes virus, which has been used to effect good as a biological control agent in other regions. A survey of adult beetles was carried out throughout Malaysia using pheromone traps. Captured beetles were examined for presence of virus using both visual/microscopic examination and PCR detection methods. The survey indicated that Oryctes virus was common in Malaysia among the adult beetles. Viral DNA analysis was carried out after restriction with HindIII enzyme and indicated at least three distinct viral genotypes. Bioassays were used to compare the viral strains and demonstrate that one strain (type B) is the most virulent against both larvae and adults of the beetle. Virus type B has been cultured and released into healthy populations where another strain (type A) forms the natural background. Capture and examination of beetles from the release site and surrounding area has shown that the spread and persistence of the applied virus strain is accompanied by a reduction in palm frond damage.     

Ecological methods for adult populations of Oryctes rhinoceros (Coleoptera, Scarabaeidae)

ABSTRACT. 1. For comparing assessment methods, O.rhinoceros (L.) populations were monitored in five 30–50 ha plots in southern Luzon, Philippines. No consistent correlations were found between number of beetles caught in traps, amount of palm damage and number of breeding sites. Apparently, plots were too small to account for fast dispersal of beetles.
2. Coconut cap traps baited with ethylchrysanthemumate attracted O.rhinoceros adults searching for breeding sites. Several factors were identified influencing catches but trapping was found to be unsuitable for accurately assessing pest populations.
3. A new method which gives estimates on the monthly number of O.rhinoceros attacks on coconut palms, was tested in the Maldives. Palms are climbed about once a year and the sequence of fronds, the number and the position of beetle cuts are recorded.
4. Reduction with age of the length of three teeth on the fore tibia of O. rhinoceros adults was studied in the Philippines. The data was insufficient to obtain a clear correlation between age and length of teeth.
5. The observations indicated that in the Philippines after leaving their site of pupation, O.rhinoceros adults spend about 5 weeks feeding on coconut palms. This is followed by a period of about 7 weeks in breeding sites and, on occasion, additional visits to palms. With these estimates it was possible to relate palm damage records with numbers of feeding adults.     

Establishment of a loop‐mediated isothermal amplification system for on‐site diagnosis of Oryctes rhinoceros nudivirus in Allomyrina dichotoma (Coleoptera: Scarabaeidae)

The Korean insect industry is rapidly developing, with the Korean horn beetle Allomyrina dichotoma as one of the most popular insects kept as pets. Korean horn beetles reared in local farms are suffering from Oryctes rhinoceros nudivirus (OrNV). In a nationwide investigation on the early death of young larvae due to this virus, 70 to 77% mortality was found. It was also confirmed that several larvae collected from some wild horn beetle habitats were infected with the virus. Thus, it was concluded that the virus could be transmitted vertically from an infected adult to the offspring and that the OrNV disease is a very serious threat to the wild horn beetle in Korea. With local farmers expecting that an early detection technique would be helpful for the quick removal of infected larvae, an on‐site diagnosis method for the viral disease using loop‐mediated isothermal amplification (LAMP) assay was thus tested. To avoid the DNA extraction process, the LAMP assay used a 50‐fold diluted hemolymph was set for diagnosis standardization and convenient on‐site application to infected larvae from local farms. A. dichotoma larvae were assayed for the OrNV infectivity with LAMP primers targeting the OrNV_gp102 gene. To evaluate the LAMP specificity, two bacterial pathogens, Bacillus thuringiensis and Serratia marcescens, causing disease in A. dichotoma were tested along with OrNV. The positive results were detected only from the OrNV‐infected larvae.     

Conserving the African rhinoceros

The Southern African rhinoceros population has been dwindling through the years. A lot of effort has been put toward coming up with the most effective way of protecting the black and white rhinoceros population in the world yet still the animals remain endangered despite the enormous global efforts to protect the species. Such conservation efforts include; establishment of sanctuaries, wildlife farming, dehorning, trade and market controls, listing of species as endangered, use of biotechnology and biotelemetry. However useful, each of these methods has its own strengths and weaknesses relative to the success of rhinoceros conservation. It however has been noted in this review that dehorning is the most effective method as it involves removal of the horn hence extinguishing the need for poaching. Despite the high success rate it has also been noted that this requires a great deal of skill and finance hence limiting the application of this technique in developing nations. Sanctuaries provide an ideal breeding environment that is better than the wild and enhanced breeding accounted for a significant increase in rhinoceros population where it has been applied. It was however been conclude that a holistic conservation approach is required to conserve the African rhinoceros.     

Social media posts suggest that coconut rhinoceros beetle has established in the Western hemisphere

The coconut rhinoceros beetle (CRB: Oryctes rhinoceros L.) is one of the most damaging pests of coconut and oil palms in the Asia/Pacific region but has not been reported from the Western Hemisphere. Here, we report a possible establishment of CRB in Nayarit, Mexico, based on an analysis of social media reports and photographs indicating the presence of the beetle and the distinctive v-shaped notches in palm fronds caused by the beetle feeding. CRB populations are destructive once established, therefore, urgent actions are required to contain the pest and prevent damage to coconut and oil palm in the Americas.     

Introducing Baculovirus oryctes Huger into Oryctes monoceros Oliv. populations in Tanzania

A follow-up of a Baculovirus oryctes release into the population of Oryctes monoceros in coconut plantations in Tanzania has been made to clarify important points which were not fully considered in an earlier publication. The low recovery of virus-infected field-collected beetles (29%) suggested that the release of B. oryctes virus was not a suitable method for the control of O. monoceros. The relatively dry conditions in the coconut growing areas were partly contributory since the virus is non-occluded. No significant and sustainable reduction in frond damage was obtained after the virus had established itself in the natural O. monoceros population. The status of O. monoceros as a serious pest of coconut is also revised since the beetle affects young plantations more than older plantations and control methods can be directed at picking up feeding insects by hooking and disposal of breeding sites.     

Surface Chemicals Inform about Sex and Breeding Status in the Biparental Burying Beetle Nicrophorus vespilloides

The multicomponent nature of chemical cues and signals are not very well understood. One reason for the often found complexity of chemical blends might be that they provide multiple messages. Burying beetles which use vertebrate carcasses as food for their larvae and defend these carcasses against inter- and intraspecific competitors are able to recognise the sex and breeding status of conspecifics. Studies have shown that the chemical composition of cuticular lipids is correlated with sex and breeding status, but there is no definitive evidence that these chemicals function in recognition. In the present study, we performed behavioural bioassays to directly asses the role of chemical cues in the recognition system of the burying beetle Nicrophorus vespilloides . After finding a carcass, females were more tolerant of dead males than of females. The behaviour was reversed when a solvent extract from the opposite sex was applied. An earlier experiment had shown that females breeding on a carcass treat non-breeding males more aggressively than breeding ones. In the present study, we could trigger the same dichotomous behaviours by presenting a single elytron from a breeding and a non-breeding beetle. In an additional experiment, females tolerated the elytra of non-breeding beetles if we had first applied an extract from a breeding beetle to these elytra. Our study is the first behavioural proof that female burying beetles obtain multiple information from chemical cues.     

Behavioural ecology of the Greater one-horned rhinoceros (Rhinoceros unicornis)

In the Chitawan Valley of Nepal there were estimated to be between 270 and 310 Greater one-horned or Indian rhinoceroses (Rhinoceros unicornis L.). Population densities reached 4.85 rhinos/km² in the favoured areas of high diversity of early successional vegetation types on the valley floor. The overall population composition was 32.3% adult females, 19.9% adult males, 21.2% sub-adults and 26.6% calves. Females first calved at a mean age of 7.1 years and the median intercalving interval was 2.8 years. Causes of death included poaching, tiger predation on calves and fighting among males. The population was increasing.
Rhinos fed from 183 species of plants belonging to 57 botanical families but grass (50 species) made up between 70 and 89% of their diet according to the season. Considerable seasonal variations in food availability resulted in movements of rhinos between vegetation types. Rhinos' ranges were smallest in the areas of greatest vegetational diversity.
Rhinos rarely formed groups. The most common type consisted of sub-adults–mainly males. Ten auditory displays were distinguished and visual displays, although less striking, included baring the lower incisor tusks. Scents were carried in the dung, the urine and the pedal scent glands. Squirt urination and foot-dragging displays were performed by breeding males only.
There was some degree of range exclusivity among breeding males but no true territoriality. Poor visibility and the relatively unpredictable distribution of resources in time and space have perhaps selected against a territorial mating system. Relationships between ecology and social organization are discussed with reference to other rhino species.
Threats to the continued survival of the Indian rhino include poaching, agricultural encroachment and erosion. In order to spread the risk of a catastrophe, reintroductions of rhinos to other protected areas are proposed.     

Studies on Rhabdionvirus oryctes: III. Incidence in the Oryctes rhinoceros population of Western Samoa

The viral pathogen Rhabdionvirus oryctes [ROV] was found to be widespread in the population of Oryctes rhinoceros in Western Samoa. About 3% of the larvae and 35% of the adults were infected with ROV. Larval breeding sites more frequently contained ROV-infected insects if the sites were occupied by two broods instead of one and if there was a high density of breeding in the area. Females infected with ROV visited the breeding sites less frequently for egg-laying than healthy females. The male adults, as they increased in age, were found to be more frequently infected with ROV, but the young female adults were more frequently infected than very young or old females. The ROV-infections in adults apparently play a more important role in suppressing the O. rhinoceros population than those of larvae.     

薇甘菊甲醇提取物对二疣犀甲生长发育的影响

【目的】探讨入侵杂草薇甘菊Mikania micrantha的利用价值, 及其在棕榈害虫二疣犀甲Oryctes rhinoceros生态防控中的应用前景。【方法】采用浸渍法和室内饲喂法, 研究了薇甘菊甲醇提取物对二疣犀甲取食量、 卵孵化、 化蛹、 羽化及幼虫发育等的影响。【结果】薇甘菊提取物对二疣犀甲具有很好的生长发育调节活性。拒食活性研究结果表明, 在不同供试浓度下, 其对二疣犀甲3龄幼虫均表现拒食活性, 且拒食率与处理浓度呈正相关。薇甘菊提取物处理二疣犀甲的卵后, 孵化率明显降低, 且孵化期延长, 10 mg/mL提取物处理后孵化率仅达66.66%, 孵化期比对照延迟3 d, 同时初孵幼虫死亡率也高达40.43%。采用添加薇甘菊提取物的饲料饲喂1龄幼虫后, 幼虫体重增长减缓, 在处理浓度为10和5 mg/g时, 处理后90 d体重增加量分别为3.83 g和4.53 g, 而对照组体重增加量达到6.87 g。薇甘菊提取物处理老熟幼虫后, 对其化蛹具有抑制作用, 造成化蛹率降低, 化蛹时间延长及蛹的畸形。经薇甘菊提取物处理二疣犀甲蛹后, 成虫羽化率降低, 羽化时间明显延长, 且畸形成虫数量增加, 主要表现为翅无法正常伸展、 虫体瘦小、 无法正常爬行等。【结论】薇甘菊具有开发为生物源昆虫生长调节剂、 并在二疣犀甲防控中应用的潜力。     

An Opportunistic Bacterial Pathogen, Pseudomonas alcaligenes , May Limit the Perpetuation of Oryctes Virus, a Biocontrol Agent of Oryctes rhinoceros L.

Pseudomonas alcaligenes was detected at a high concentration (10 9 -10 10 cells mL -1 ) in the haemolymph of some dead Oryctes rhinoceros grubs collected from its breeding sites in the three southern districts, viz. Alleppey, Quilon and Kottayam of Kerala State, India. In a laboratory colony maintained for production of Oryctes virus, an important biocontrol agent of this major coconut pest, approximately 52% of the grubs succumbed to septicaemia with similar symptoms. The bacterium was found to be a component of the gut microflora of healthy grubs. Occurrence of the viral infection naturally or when induced in the laboratory in the O. rhinoceros grubs, appeared to be one of the biotic stress factors for P.alcaligenes to become an opportunistic pathogen. A preponderance of this bacterial infection in field populations during the periods when natural viral infection in grubs was above average, agrees with this observation. This finding becomes significant as infection by the opportunistic bacterial pathogen, P. alcaligenes , reduces the production of Oryctes virus inoculum in nature and limits the field-perpetuation of this viral biocontrol agent.     

Variation in cross‐sectional horn shape within and among rhinoceros beetle species

Sexual selection has equipped male rhinoceros beetles with large horns on their head and prothorax to aid in battle over access to females. Horns are used to pry and dislodge opponents from resource sites that attract females, so an optimal horn should be able both to withstand the high stresses imposed during fights, and to resist deflection in response to these loads. We examined the cross‐sectional morphology of horns using micro‐computed tomography scanning to determine how horn structure changes with horn length to withstand the different fighting loads. Specifically, we measured the second moment of area of horns within and among rhinoceros beetle species to assess whether changes in cross‐sectional morphology accompany changes in body size in order to maintain high strength and stiffness during fights. We find that the second moment of area of horns increases with body size both intra‐specifically and inter‐specifically, and that these relationships closely fit those predicted if horns have been selected to be strong and stiff fighting structures. Our results therefore support the hypothesis that rhinoceros beetle horns are structurally adapted for combat.     

The smell of parents: breeding status influences cuticular hydrocarbon pattern in the burying beetle Nicrophorus vespilloides

The waxy layer of the cuticle has been shown to play a fundamental role in recognition systems of insects. The biparental burying beetle Nicrophorus vespilloides is known to have the ability to discriminate between breeding and non-breeding conspecifics and also here cuticular substances could function as recognition cue. However, it has not yet been demonstrated that the pattern of cuticular lipids can reflect the breeding status of a beetle or of any other insect. With chemical analysis using coupled gas chromatography-mass spectrometry, we showed that the chemical signature of N. vespilloides males and females is highly complex and changes its feature with breeding status. Parental beetles were characterized by a higher amount of some unusual unsaturated hydrocarbons than beetles which are not caring for larvae. The striking correlation between cuticular profiles and breeding status suggests that cuticular hydrocarbons inform the beetles about parental state and thus enable them to discriminate between their breeding partner and a conspecific intruder. Furthermore, we found evidence that nutritional conditions also influence the cuticular profile and discuss the possibility that the diet provides the precursors for the unsaturated hydrocarbons observed in parental beetles. Our study underlines the fact that the cuticular pattern is rich of information and plays a central role in the burying beetles' communication systems.     

The role of doublesex in the evolution of exaggerated horns in the Japanese rhinoceros beetle

Male‐specific exaggerated horns are an evolutionary novelty and have diverged rapidly via intrasexual selection. Here, we investigated the function of the conserved sex‐determination gene doublesex (dsx) in the Japanese rhinoceros beetle (Trypoxylus dichotomus) using RNA interference (RNAi). Our results show that the sex‐specific T. dichotomus dsx isoforms have an antagonistic function for head horn formation and only the male isoform has a role for thoracic horn formation. These results indicate that the novel sex‐specific regulation of dsx during horn morphogenesis might have been the key evolutionary developmental event at the transition from sexually monomorphic to sexually dimorphic horns.